With the widespread use of yeast two-hybrid systems, many heterodimeric forms of S100 proteins have been found, although their biological significance is unknown. In the present study, S100A1 was found to interact with another S100 protein, S100P, by using the yeast two-hybrid system. The binding parameters of the interaction were obtained using an optical biosensor and show that S100P has a slightly higher affinity for S100A1 (Kd=10–20 nM) when compared with that for self-association (Kd=40–120 nM). The physical interaction of S100A1 and S100P was also demonstrated in living mammalian cells using a fluorescence resonance energy transfer technique. Preincubation of recombinant S100P with S100A1, before the biosensor assay, reduced by up to 50% the binding of S100P to a recombinant C-terminal fragment of non-muscle myosin A, one of its target molecules. Site-specific mutations of S100P and S100A1, combined with homology modelling of an S100P/S100A1 heterodimer using known S100P and S100A1 structures, allowed the hydrophobic interactions at the dimeric interface of the heterodimer to be defined and provide an explanation for the heterodimerization of S100P and S100A1 at the molecular level. These results have revealed the similarities and the differences between the S100P homodimer and the S100A1/S100P heterodimer.
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Research Article|
August 10 2004
Heterodimeric interaction and interfaces of S100A1 and S100P
Guozheng WANG;
Guozheng WANG
1
*Cancer and Polio Research Fund Laboratories Molecular Medicine Group, School of Biological Sciences, Biosciences Building, University of Liverpool, Crown Street, Liverpool L69 7ZB, U.K.
1To whom correspondence should be addressed (email wangg@liv.ac.uk).
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Shu ZHANG;
Shu ZHANG
*Cancer and Polio Research Fund Laboratories Molecular Medicine Group, School of Biological Sciences, Biosciences Building, University of Liverpool, Crown Street, Liverpool L69 7ZB, U.K.
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David G. FERNIG;
David G. FERNIG
*Cancer and Polio Research Fund Laboratories Molecular Medicine Group, School of Biological Sciences, Biosciences Building, University of Liverpool, Crown Street, Liverpool L69 7ZB, U.K.
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David SPILLER;
David SPILLER
†Molecular Imaging Laboratory, School of Biological Sciences, Biosciences Building, University of Liverpool, Crown Street, Liverpool L69 7ZB, U.K.
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Marisa MARTIN-FERNANDEZ;
Marisa MARTIN-FERNANDEZ
‡CLRC Daresbury Laboratory, Warrington WA4 4AD, U.K.
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Hongmei ZHANG;
Hongmei ZHANG
§Laboratory of Structural Biology and Ministry of Education Laboratory of Protein Science, School of Life Science and Engineering, Tsinghua University, Beijing 100084, People's Republic of China
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Yi DING;
Yi DING
§Laboratory of Structural Biology and Ministry of Education Laboratory of Protein Science, School of Life Science and Engineering, Tsinghua University, Beijing 100084, People's Republic of China
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Zihe RAO;
Zihe RAO
§Laboratory of Structural Biology and Ministry of Education Laboratory of Protein Science, School of Life Science and Engineering, Tsinghua University, Beijing 100084, People's Republic of China
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Philip S. RUDLAND;
Philip S. RUDLAND
*Cancer and Polio Research Fund Laboratories Molecular Medicine Group, School of Biological Sciences, Biosciences Building, University of Liverpool, Crown Street, Liverpool L69 7ZB, U.K.
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Roger BARRACLOUGH
Roger BARRACLOUGH
*Cancer and Polio Research Fund Laboratories Molecular Medicine Group, School of Biological Sciences, Biosciences Building, University of Liverpool, Crown Street, Liverpool L69 7ZB, U.K.
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Publisher: Portland Press Ltd
Received:
January 26 2004
Revision Received:
May 24 2004
Accepted:
June 01 2004
Accepted Manuscript online:
June 01 2004
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London
2004
Biochem J (2004) 382 (1): 375–383.
Article history
Received:
January 26 2004
Revision Received:
May 24 2004
Accepted:
June 01 2004
Accepted Manuscript online:
June 01 2004
Citation
Guozheng WANG, Shu ZHANG, David G. FERNIG, David SPILLER, Marisa MARTIN-FERNANDEZ, Hongmei ZHANG, Yi DING, Zihe RAO, Philip S. RUDLAND, Roger BARRACLOUGH; Heterodimeric interaction and interfaces of S100A1 and S100P. Biochem J 15 August 2004; 382 (1): 375–383. doi: https://doi.org/10.1042/BJ20040142
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